Professional Documents
Culture Documents
Made by :
Adel Mohie
Under supervision of
Dr:Sofia Khalil
What are restriction enzymes?
Blunt ends
Sticky ends
Blunt ends
• Some restriction enzymes cut DNA at opposite base
• They leave blunt ended DNA fragments
• These blunt ended fragments can be joined to any other
DNA fragment with blunt ends.
• Enzymes useful for certain types of DNA cloning experiments
.
Sticky ends
• Most restriction enzymes make staggered
cuts
• Staggered cuts produce single stranded
“sticky-ends”
“Sticky Ends” Are Useful
E Escherichia genus
co coli species
R RY13 strain
order of identification
I First identified
in the bacterium
TYPES OF RESTRICTION ENZYMES
• Type I
• Type II
• Type III
• These types are categorization based on:
Their composition.
Enzyme co-factor requirement.
the nature of their target sequence.
position of their DNA cleavage site relative to
the target sequence.
Type I
• Capable of both restriction and modification
activities
• The co factors S-Adenosyl Methionine(AdoMet), ATP,
and mg+are required for their full activity
• Contain:
two R(restriction) subunits
two M(methylation) subunits
one S(specifity) subunits
• Cleave DNA at random length from recognition sites
Type II
• These are the most commonly available and used
restriction enzymes
• They are composed of only one subunit.
• Their recognition sites are usually undivided and
palindromic and 4-8 nucleotides in length,
• they recognize and cleave DNA at the same site.
• They do not use ATP for their activity
• they usually require only Mg2+ as a cofactor.
Type III
• Type III restriction enzymes ) recognize two separate
non-palindromic sequences that are inversely
oriented.
• They cut DNA about 20-30 base pairs after the
recognition site.
• These enzymes contain more than one subunit.
• And require AdoMet and ATP cofactors for their roles
in DNA methylation and restriction
Type IV
• Cleave only normal and modified DNA
(methylated, hydroxymethylated and glucosyl-
hydroxymethylated bases).
• Recognition sequences have not been well
defined
• Cleavage takes place ~30 bp away from one of
the sites
APPLICATION OF RESTRICTION ENZYMES